Molecular identification of human/goat xenogeneic model.
- Author:
Meijue CHEN
1
;
Jingbin YAN
;
Yudan FANG
;
Zhaorui REN
;
Yanping XIAO
;
Shuzhen HUANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Antigens, CD34; genetics; Blotting, Southern; Female; Genes, sry; genetics; Glycophorin; genetics; Goats; Hematopoietic Stem Cell Transplantation; methods; Hematopoietic Stem Cells; cytology; metabolism; Humans; In Situ Hybridization, Fluorescence; Male; Polymerase Chain Reaction; Transplantation Chimera; genetics; Transplantation, Heterologous
- From: Chinese Journal of Hematology 2002;23(12):634-637
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo identify the human hematopoietic stem cells from the human/goat xenogeneic model with molecular techniques.
METHODSDNA and total RNA were extracted from 11 transplanted goat peripheral blood cells. Human CD(34), GPA and SRY genes were amplified with PCR in these samples, and CD(34), GPA mRNA transcripts were detected using RT-PCR in 5 and 6 goat peripheral blood cells, respectively. Southern blot analysis was performed in 8 goat DNAs to detect the human specific alpha-satellite sequence. Meanwhile FISH was also performed to detect the human cells in goat blood with a probe of human Y chromosome.
RESULTSHuman CD(34) and GPA genes could be detected with PCR in all the 11 goats, and SRY gene did in 5 goats transplanted with hematopoietic stem cells derived from male human babies. Southern blot showed that human specific alpha-satellite sequence was present in 8 goats. By RT-PCR, human CD(34) mRNA was detected in 5 experimental goats, GPA mRNA was found in the other 6 experimental goats and FISH assay showed that some peripheral blood cells of the human/goat xenogeneic model were positive.
CONCLUSIONExistence of human cells in the recipient goats was identified by molecular detection, which was feasible for the examination of human/goat xenogeneic models.
